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Lookup NU author(s): Dr Ian Cowell,
Professor Caroline AustinORCiD
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© 2018, Springer Science+Business Media, LLC.The TARDIS assay was originally developed as a means of detecting and quantifying melphalan and cisplatin DNA adducts at the single cell level, but it has since been adapted to quantify topoisomerase DNA complexes that result from the actions of topoisomerase poisons and this is currently the main use of the assay. The method employs sensitive immunofluorescent detection to quantify topoisomerase molecules covalently coupled to DNA in what are often referred to as cleavage complexes. Free topoisomerase molecules, and other cellular constituents are first removed using salt-detergent extraction of agarose-embedded, unfixed cells. Using these stringent extraction conditions, genomic DNA remains in place in the agarose as “nuclear ghosts,” and any covalent attached molecules can be detected and quantified by immunofluorescence with a low background.
Author(s): Cowell IG, Austin CA
Publication type: Book Chapter
Publication status: Published
Book Title: DNA Topoisomerases: Methods and Protocols
Online publication date: 25/11/2017
Acceptance date: 02/04/2016
Series Title: Methods in Molecular Biology
Publisher: Humana Press
Place Published: New York
Library holdings: Search Newcastle University Library for this item